CN103667053A - Cell culture device capable of automatically and continuously replacing cell culture fluid - Google Patents

Cell culture device capable of automatically and continuously replacing cell culture fluid Download PDF

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Publication number
CN103667053A
CN103667053A CN201310182185.8A CN201310182185A CN103667053A CN 103667053 A CN103667053 A CN 103667053A CN 201310182185 A CN201310182185 A CN 201310182185A CN 103667053 A CN103667053 A CN 103667053A
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cell culture
fluid
cover
nutrient solution
cell
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CN201310182185.8A
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CN103667053B (en
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李莹辉
谭映军
王春艳
于建茹
聂捷琳
顾寅
万玉民
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China Astronaut Research and Training Center
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China Astronaut Research and Training Center
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
    • C12M41/24—Heat exchange systems, e.g. heat jackets or outer envelopes inside the vessel
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/34—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of gas

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  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
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  • Life Sciences & Earth Sciences (AREA)
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Abstract

A cell culture device is capable of automatically and continuously replacing cell culture fluid and providing the conditions required for the cell culture, thereby realizing the automatic and continuous replacement of cell culture fluid in the culture of cell. The device is characterized by comprising an environmental control system, a continuous culture system and a shell, wherein the shell is composed of a main body structure, a right side cover, a main cover, a cover lock and a hinge; the main cover is mounted at the front surface of the main body structure by virtue of the hinge and locked on the main body structure by virtue of the cover lock; the right side cover is fixed on the right side of the main body structure by screws; the environmental control system consists of a heater, a convection fan, a temperature sensor, a thermal insulation structure, CO2 cylinders, an air delivery pipe, a solenoid valve, CO2 sensors, control circuit boards, a display screen and connecting lines; the continuous culture system comprises culture fluid storage bottles, pipelines, a fluid replacement motor, a fluid replacement pump head, fluid inlet pipes, a cell culture bottle cover, fluid output pipes, cell culture flasks, a sample holder and waste fluid collection bottles.

Description

Automatic continuous changes liquid cell culture apparatus
Technical field the present invention relates to a kind of cell culture apparatus that is applied to cytobiology field and life science, particularly a kind of cell culture apparatus that possesses automatic replacing nutrient solution function.
Background technology is in RESEARCH ON CELL-BIOLOGY field and life science field, and cell cultures is the most basic link.In cell cultivation process, changing at regular time and quantity of cell culture fluid is the operation of most critical.At present, the conventional cell culture fluid substitute mode in laboratory realizes by operation manually.It is as follows that this cell cultures replacing method is not enough:
1, artificial hand is moved liquid operation discontinuous, cannot in cell cultivation process, realize continuous and quantitative cell cultures fluid exchange.
2, artificial hand is moved liquid and cell sample need to be taken out from culture environment to operation, and the variation of environment can not be ignored the impact of cell sample.
3, manually change liquid and be difficult to precisely control nutrient solution consumption.
Summary of the invention the object of the invention is to set up a kind of Automatic continuous and changes liquid cell culture apparatus, has both possessed continuously and has certainly moved hydraulic control ability processed, can provide again cell cultures needed condition, realizes cell culture fluid automatic continuous replacing in culturing process.
The present invention includes environmental control system, continuous culture system and shell three parts, continuous culture system is installed on environmental control system inside, and environmental control system is built in shell.
Described shell is comprised of agent structure, right cap, main lid, cover lock and flap.Main lid is installed on by flap before agent structure, by cover lock, is locked in agent structure; Right cap is fixed on agent structure right side by screw.
Described environmental control system is comprised of well heater, convection fan, temperature sensor, insulation construction, CO2 gas cylinder, pneumatic tube, magnetic valve, CO2 sensor, control circuit board, display screen and tie.Insulation construction is installed on agent structure left side; Well heater and convection fan are installed on the insulation construction bottom surface upper left corner with screw, and temperature sensor is bonded at upper position in the middle of insulation construction bottom surface; It is following that magnetic valve is installed on insulation construction right hand external with screw, and CO2 sensor is bonded at middle position on the lower, insulation construction bottom surface, is positioned at temperature sensor below; Control circuit board is installed on top, outside, insulation construction right side, by screw, fixes; Display screen is arranged on right side before agent structure with screw.Well heater, convection fan, temperature sensor, magnetic valve, CO2 sensor and display screen are all connected with control circuit board by tie.External CO2 gas cylinder and magnetic valve import are connected by the pneumatic tube through the lower right corner, the agent structure back side, and electromagnetic valve outlet is connected to insulation construction inside with pneumatic tube, form the controlled flowing passage of the insulation construction inside of CO2 gas from gas cylinder to culturing cell.
Described continuous culture system by nutrient solution storage bottle, pipeline, change liquid motor, change liquid pump head, nutrient solution input tube, cell cultures bottle cap, nutrient solution output tube, Tissue Culture Flask, sample rack and waste collection bottle form.Nutrient solution memory and waste collection bottle are positioned over insulation construction right side, change liquid motor and with screw, are installed on the position of keeping right, insulation construction bottom surface with changing liquid pump head, change liquid motor and by motor shaft band, move liquid pump head and work; Tissue Culture Flask is positioned over insulation construction left side with sample rack, and cell cultures bottle cap is screwed in cell cultures bottleneck by screw thread, and nutrient solution input tube and nutrient solution output tube stretch in cell cultures after inserting cell cultures bottle cap; Pipeline connects nutrient solution storage bottle, changes liquid pump head, nutrient solution input tube, Tissue Culture Flask, nutrient solution output tube and waste collection bottle, forms a complete cell culture fluid stream and adds loop.
The present invention adopts after technique scheme, and its distinguishing feature is:
1, can in cell cultivation process, realize automated cell nutrient solution and change continuously, cell cultures flow velocity can regulate according to requirement of experiment; Also can change liquid by automatic intermittent.
2, change liquid process and culturing process and organically combine, device can be controlled envrionment temperature and the CO2 gas concentration of insulation construction inside automatically, meets cell cultures requirement, can not affect normal cell cultivation because changing liquid.Therefore, the present invention had both possessed cell automated cell and had cultivated function, possessed again and changed continuously liquid function simultaneously, and the two has both.
3, change the whole automatizations of liquid process, do not relate to manual operation, change liquid measure control accuracy high.
Description of drawings 1 structural representation of the present invention
Embodiment agent structure 25 adopts hard aluminium alloy LY12 machining global formation, and physical dimension is 541.6 * 272 * 200 (units: mm), black moulding sprayed on surface.Main lid 2 also adopts LY12 machining global formation, black the moulding of surface spray, by 2 standard flaps 3, be installed on agent structure 25 front, left side, cover lock 1 adopts stainless steel 1Cr18Ni9Ti processing, can in main lid 2 open holess, rotate, cover lock 1 rotates 90 ° and main lid 2 can be locked in agent structure 25, then rotates 90 ° and get final product release; Right cap 15 is aluminium alloy LY12 plate, by 12 M3 screws, is fixed on agent structure 25 right sides.
Insulation construction 24 adopts nontoxic PC material foaming, and net thickness is 10mm, with epoxy construction, is adhered to agent structure 25 inside left; Well heater 22 adopts JRP125 type Kapton type, convection fan 23 is standard DC12V axial fan, well heater 22 and 4 M3 screws of convection fan 23 use are installed on insulation construction 24 upper left corners, bottom surface, temperature sensor 21 adopts numeric type sensor, model is MCP9804 type, by structure glue, is bonded at middle upper position, insulation construction 24 bottom surfaces; The gas liquid two purpose valve that magnetic valve 13 is low seepage, model is NV-24 miniature electromagnetic valve, is installed on insulation construction 224 right hand external following, CO with 2 M3 screws 2sensor 20 is COZIR type near infrared sensor, is bonded at middle position on the lower, insulation construction 24 bottom surfaces, is positioned at temperature sensor 21 belows; Control circuit board is installed on insulation construction 24 tops, outside, right side with 4 M3 standard sunk screws; Display screen 12 is 8 " digital touch panel; can the direct input unit working parameter of digitizing, experimental installation enters after working order, and display screen 12 is display unit real work parameter also; display screen 12 models are AMT9507 type, with 5 M3 sunk screws, is arranged on right side before agent structure 25.External CO 2gas cylinder 16 is the flat pressure cylinder of 8L aluminium matter, and air storage pressure is 5MPa, CO 2the import of gas cylinder 16 and magnetic valve 13 is connected by the pneumatic tube 16 through agent structure 25 lower right corner, the back side, and pneumatic tube 16 materials are tetrafluoroethylene, and pipe external diameter is 3.2mm, wall thickness 1mm, and magnetic valve 13 outlets are connected to insulation construction 24 inside with pneumatic tube 16.
Nutrient solution storage bottle 11 and waste collection bottle 10 are glassiness reagent bottle with cover, nutrient solution storage bottle 11 and waste collection bottle 10 are all positioned over insulation construction 24 right sides, change liquid motor 19 for micro-stepping SSM23S type stepper-motor, change liquid pump head 18 for card form DG-12 standard Wriggling Pump Head, change 4 M4 screws of liquid motor 19 use and be installed on the position of keeping right, insulation construction 24 bottom surfaces, change liquid motor 19 and move 18 work of liquid pump head by motor shaft band; Tissue Culture Flask 5 is standard 25cm 2tissue Culture Flask, 3 Tissue Culture Flask 5 use sample racks 4 are positioned over insulation construction 24 left sides, and the quantity of Tissue Culture Flask 5 can increase and decrease with requirement of experiment; Sample rack 4 adopts aluminium alloy LY12 line cutting global formation, surface passivation place.Cell cultures bottle cap 6 is screwed in Tissue Culture Flask 5 bottlenecks by screw thread, after nutrient solution input tube 8 and nutrient solution output tube 7 insert cell cultures bottle cap 6, stretches in Tissue Culture Flask 5; Pipeline 9 is silicone rubber tube, pipe external diameter 5mm, and wall thickness is 2mm, connects nutrient solution storage bottle 11, changes liquid pump head 18, nutrient solution input tube 8, Tissue Culture Flask 5, nutrient solution output tube 7 and waste collection bottle 10, forms a complete cell culture fluid stream and adds loop.
During experiment, first press requirement of experiment by the Tissue Culture Flask of some amount 5, in super clean bench, with pipeline 9, connect according to the order of sequence nutrient solution storage bottle 11 and waste collection bottle 10, form a complete stream.Tissue Culture Flask 5 is positioned on sample rack 4, nutrient solution storage bottle 11 and waste collection bottle 10 are positioned over insulation construction 24 right sides again, and pipeline 9 is installed to and changes on liquid pump head 18.Close upper main cover 2, with cover lock 1 locking.After device energising, set temperature, CO on display screen 12 2the working parameter such as concentration and flow velocity, and startup enters working order.In working process, change liquid motor 18 bands and move 19 work of liquid pump head, cell culture fluid on request flow velocity, from nutrient solution storage bottle 11 flows out, enters in each Tissue Culture Flask 5, and the nutrient solution of outflow is collected in waste collection bottle 10 automatically; Meanwhile, device, by regulating well heater 22 and convection fan 23, is controlled the temperature of insulation construction 24 inside automatically; Electromagnetic valve for adjusting 13, controls and from gas cylinder 17, along pneumatic tube 16, enters the CO of insulation construction 24 inside 2amount, regulates CO 2concentration, thus realize when certainly moving liquid continuously, automatically carry out cell cultures.

Claims (1)

1. Automatic continuous changes a liquid cell culture apparatus, it is characterized in that: comprise environmental control system, continuous culture system and shell three parts; Described shell is comprised of agent structure, right cap, main lid, cover lock and flap, and main lid is installed on by flap before agent structure, by cover lock, is locked in agent structure, and right cap is fixed on agent structure right side by screw; Described environmental control system is by well heater, convection fan, temperature sensor, insulation construction, CO 2gas cylinder, pneumatic tube, magnetic valve, CO 2sensor, control circuit board, display screen and tie form; Described continuous culture system by nutrient solution storage bottle, pipeline, change liquid motor, change liquid pump head, nutrient solution input tube, cell cultures bottle cap, nutrient solution output tube, Tissue Culture Flask, sample rack and waste collection bottle form.
CN201310182185.8A 2013-05-17 2013-05-17 Automatic continuous changes liquid cell culture apparatus Active CN103667053B (en)

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104371920A (en) * 2014-11-06 2015-02-25 徐小杨 Liquid drop exchange operating device for cell culture
CN107585876A (en) * 2017-08-30 2018-01-16 宁波佗鹊堂生物科技有限公司 The material-changing method of mother liquor culture
CN107881107A (en) * 2017-11-15 2018-04-06 中国航天员科研训练中心 The image capturing system of automated cell culture device
CN109294914A (en) * 2018-11-19 2019-02-01 冯可 One kind can remotely monitor multi-functional culture apparatus
CN109735453A (en) * 2019-02-26 2019-05-10 陕西师范大学 A kind of breast cancer cell line MCF-7 at the uniform velocity changes liquid culture apparatus
CN110951612A (en) * 2019-12-12 2020-04-03 中国科学院深圳先进技术研究院 Automatic liquid changing device

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006141326A (en) * 2004-11-22 2006-06-08 Hitachi Medical Corp Culturing device
CN101993823A (en) * 2009-08-28 2011-03-30 中国航天员科研训练中心 Environmental self-control type cytology experimental platform
CN102382766A (en) * 2011-11-29 2012-03-21 向开兴 Tidal bioreactor for animal cell culture
CN102533548A (en) * 2010-12-22 2012-07-04 株式会社日立制作所 Cell culture apparatus
JP2013031461A (en) * 2012-11-16 2013-02-14 Hitachi Ltd Automated culturing apparatus

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006141326A (en) * 2004-11-22 2006-06-08 Hitachi Medical Corp Culturing device
CN101993823A (en) * 2009-08-28 2011-03-30 中国航天员科研训练中心 Environmental self-control type cytology experimental platform
CN102533548A (en) * 2010-12-22 2012-07-04 株式会社日立制作所 Cell culture apparatus
CN102382766A (en) * 2011-11-29 2012-03-21 向开兴 Tidal bioreactor for animal cell culture
JP2013031461A (en) * 2012-11-16 2013-02-14 Hitachi Ltd Automated culturing apparatus

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104371920A (en) * 2014-11-06 2015-02-25 徐小杨 Liquid drop exchange operating device for cell culture
CN107585876A (en) * 2017-08-30 2018-01-16 宁波佗鹊堂生物科技有限公司 The material-changing method of mother liquor culture
CN107881107A (en) * 2017-11-15 2018-04-06 中国航天员科研训练中心 The image capturing system of automated cell culture device
CN107881107B (en) * 2017-11-15 2023-06-06 中国航天员科研训练中心 Image acquisition system of automatic cell culture equipment
CN109294914A (en) * 2018-11-19 2019-02-01 冯可 One kind can remotely monitor multi-functional culture apparatus
CN109735453A (en) * 2019-02-26 2019-05-10 陕西师范大学 A kind of breast cancer cell line MCF-7 at the uniform velocity changes liquid culture apparatus
CN110951612A (en) * 2019-12-12 2020-04-03 中国科学院深圳先进技术研究院 Automatic liquid changing device

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